Single-dose administration of factor VIIa

ABSTRACT

The present invention provides methods for preventing and/or treating bleeding episodes by administering a single dose of Factor VIIa or a Factor VIIa equivalent. Preferably, the single dose comprises between about 150 and about 500 ug/kg Factor VIIa or Factor VIIa equivalent.

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority under 35 U.S.C. 119 of U.S.application Ser. No. 60/305,720 filed on Jul. 16, 2001, the contents ofwhich are fully incorporated herein by reference.

FIELD OF THE INVENTION

[0002] The present invention relates to methods for preventing and/ortreating bleeding using coagulation factors.

BACKGROUND OF THE INVENTION

[0003] Factor VII is a plasma coagulation factor, which, once activatedto Factor VIIa, initiates the normal haemostatic process by forming acomplex with tissue factor (TF), a cell surface glycoprotein that isexposed to the circulation as a result of injury to the vessel wall.Subsequently, the Factor VIIa-TF complex activates Factor IX and FactorX into their activated forms (Factor IXa and Factor Xa, respectively).Factor Xa converts limited amounts of prothrombin to thrombin on thetissue factor-bearing cell. Thrombin activates platelets and Factors Vand VIII into Factors Va and VIIIa, both cofactors in the furtherprocess leading to the full thrombin burst. Thrombin finally convertsfibrinogen to fibrin resulting in formation of a fibrin clot. Fibrinclots formed in the presence of high thrombin concentrations comprise atighter network and are more resistant to proteolysis than clots formedin lower concentrations of thrombin. Accordingly, a full thrombin burstis likely to be important for forming a hemostatic plug that isresistant to fibrinolysis and thus to facilitate full hemostasis andwound healing.

[0004] Factor VIIa, as well as Factor VIII and Factor IX, have been usedto control bleeding disorders that are caused by clotting factordeficiencies (such as, e.g. haemophilia A and B or deficiency ofcoagulation Factors XI or VII) or clotting factor inhibitors. FactorVIIa has also been used to control excessive bleeding caused bydefective platelet function, thrombocytopenia or von Willebrand'sdisease.

[0005] Typically, however, patients are treated with multiple injectionsor infusions of a coagulation factor before the bleeding is stopped. Inthe case of Factor VIII and Factor IX administration, a considerablenumber of injections are needed to maintain haemostasis until the injurycausing the bleeding is completely healed. A quicker and more effectivetreatment, as well as a reduction in the number of injections neededbefore the bleeding is stopped, represent important benefit to suchpatients. It would also be a considerable benefit to a patient needingfrequent injections or infusions with a haemostatic agent that theinjection frequency be reduced.

[0006] Thus, there is a need in the art for methods for preventingand/or treating bleeding episodes that reduce the duration ofadministration and provide a more rapid hemostasis.

SUMMARY OF THE INVENTION

[0007] The present invention relates to methods for preventing and/ortreating a bleeding episode in a subject in need of such treatment,which are carried out by administering to the subject, in a single dose,a single-dose-effective amount of Factor VIIa or a Factor VIIaequivalent. Preferably, subsequent to the administration, no furtherFactor VIIa or protein having Factor VIIa coagulant activity isadministered to the subject for an interval of at least about 1 hour. Insome embodiments, the interval is at least about 4 hours; in otherembodiments, the interval is at least about 24 hours; and in someembodiments, no further Factor VIIa or protein having Factor VIIacoagulant activity is administered during the particular bleedingepisode that is being treated.

[0008] In some embodiments, the single-dose-effective amount comprisesbetween about 150 and about 500 ug/kg Factor VIIa or a correspondingamount of a Factor VIIa equivalent; in other embodiments, thesingle-dose-effective amount comprises between about 200 and about 500ug/kg; between about 250 and about 500 ug/kg; between about 300 andabout 500 ug/kg; between about 350 and 500 ug/kg; between about 400 andabout 500 ug/kg; between about 450 and about 500 ug/kg; and greater than500 ug/kg, respectively, of Factor VIIa or a corresponding amount of aFactor VIIa equivalent.

[0009] In some embodiments, the Factor VIIa equivalent exhibits at leastabout 30% of the coagulant activity of Factor VIIa on a molar basis.Non-limiting examples of a Factor VIIa equivalent include S52A-FVII,S60A-FVII; L305V-FVII, L305V/M306D/D309S-FVII, L3051-FVII, L305T-FVII,F374P-FVII, V158T/M298Q-FVII, V158D/E296V/M298Q-FVII K337A-FVII,M298Q-FVII, V158D/M298Q-FVII, L305V/K337A-FVII, V158D/E296V/M298Q/L305V-FVII, V158D/E296V/M298Q/K337A-FVII,V158D/E296V/M298Q/L305V/K337A-FVII, K157A-FVII, E296V-FVII,E296V/M298Q-FVII, V158D/E296V-FVII, V158D/M298K-FVII, and S336G-FVII;Factor VIIa variants exhibiting increased proteolytic stability asdisclosed in U.S. Pat. No. 5,580,560; Factor VIIa that has beenproteolytically cleaved between residues 290 and 291 or between residues315 and 316; oxidized forms of Factor VIIa; Factor VII-sequence variantswherein the amino acid residue in positions 290 and/or 291 (of SEQ IDNO:1), preferably 290, have been replaced, and Factor VII-sequencevariants wherein the amino acid residue in positions 315 and/or 316 (ofSEQ ID NO:1), preferably 315, have been replaced.

[0010] In some embodiments, the method further comprises administering,with or substantially simultaneously with the single dose, a secondcoagulant agent. Non-limiting examples of a second coagulant agentinclude Factor VIII, Factor IX, and Factor XIII.

[0011] In some embodiments, the invention provides a method for treatinga bleeding episode, which is carried out by administering to a subjectin need of such treatment (i) a first amount of Factor VIIa or a FactorVIIa equivalent and (ii) a second amount of second coagulant agent,wherein the first and second amounts together comprise an aggregateeffective amount for treating the bleeding episode and the aggregateeffective amount is administered in a single dose.

[0012] In some embodiments, the Factor VIIa used in practicing theinvention is recombinant human Factor VIIa.

[0013] In one embodiment, the invention provides a method for treating ableeding episode, which is carried out by administering to a humansubject in need of such treatment an effective amount for treating saidbleeding of human Factor VIIa or a human Factor VIIa equivalent,wherein:

[0014] (i) said effective amount is administered in a single dose over aperiod of less than about 5 minutes;

[0015] (ii) said effective amount comprises between about 300 and about500 ug/kg human Factor VIIa or human Factor VIIa equivalent or acorresponding amount of a Factor VIIa equivalent; and

[0016] (iii) subsequent to said administration, no further Factor VIIaor Factor VIIa equivalent is administered to said subject for a periodof at least about 1 hour.

[0017] In practicing the present invention, administration may beachieved by any mode of administration, including, without limitation,intravenous, intramuscular, subcutaneous, mucosal, and pulmonary routesof administration.

[0018] In another aspect, the invention provides a method for preventinga bleeding episode, which is carried out by administering to a humansubject in need of such prevention an effective amount for preventingthe bleeding episode of human Factor VIIa or a human Factor VIIaequivalent, wherein:

[0019] (i) the effective amount is administered in a single dose over aperiod of less than about 5 minutes;

[0020] (ii) the effective amount comprises between about 250 and about500 ug/kg human Factor VIIa or human Factor VIIa equivalent or acorresponding amount of a Factor VIIa equivalent; and

[0021] (iii) subsequent to the administration, no further Factor VIIa orFactor VIIa equivalent is administered to said subject for a period ofat least about 1 hour.

[0022] In some embodiments, the subject suffers from hemophilia A or B.

[0023] In some embodiments, the bleeding is joint bleeding.

[0024] In some embodiments, the subject has not been treatedtherapeutically with an anticoagulant for at least about 48 hours priorto administration of Factor VIIa or a Factor VIIa equivalent.

[0025] In some embodiments, the subject has not been treatedtherapeutically with a Vitamin K antagonist for at least about 48 hoursprior to said administering.

DETAILED DESCRIPTION OF THE INVENTION

[0026] The present invention relates to methods for preventing and/ortreating bleeding in animals, particularly in humans. The invention isbased on the discovery that administration of a single dose comprising apredetermined amount of Factor VIIa is effective in treating bleedingepisodes, including major bleeding episodes, and can also be used toprevent an anticipated bleeding episode.

[0027] Without wishing to be bound by theory, it is believed that FactorVIIa enhances thrombin generation on tissue factor-bearing cells and onactivated platelets at the site of injury and that administration of asingle dose of Factor VIIa according to the present invention provides afull thrombin burst, thereby facilitating the formation of a tight,strong fibrin network that is relatively resistant to prematurefibrinolysis and obviating the need for further administration of FactorVIIa.

[0028] As used herein, prevention refers to prophylactic administrationof Factor VIIa so as to minimize or inhibit an anticipated bleedingepisode, such as, e.g., prior to surgery. Treatment refers to regulationof an already-occurring bleeding, such as, for example, in trauma, withthe purpose of inhibiting or minimizing the bleeding. It will beunderstood that efficacy in prevention and/or treatment according to thepresent invention encompasses the absence of significant side-effects,including, without limitation, disseminated intravascular coagulation(DIC), that would counterindicate to those of ordinary skill in the artthe use of any particular therapeutic regimen.

[0029] Bleeding refers to extravasation of blood from any component ofthe circulatory system. The term “bleeding episode” includes, withoutlimitation, bleeding (including, without limitation, excessive,uncontrolled bleeding, i.e., haemorrhaging) in connection with surgeryor trauma, such as, for example, in connection with acute haemarthroses(bleedings in joints), chronic haemophilic arthropathy, haematomas,(e.g., muscular, retroperitoneal, sublingual and retropharyngeal),bleedings in other tissue, haematuria (bleeding from the renal tract),cerebral haemorrhage, surgery (e.g., hepatectomy), dental extraction,and gastrointestinal bleedings (e.g., UGI bleeds). Also included arehemorrhages in organs such as the brain, inner ear region and eyes withlimited possibility for surgical haemostasis; as well as hemorrhages inconnection with biopsies of various organs (liver, lung, tumour tissue,gastrointestinal tract) as well as laparoscopic surgery. Common to thesesituations is the difficulty in providing haemostasis using surgicaltechniques (such as, e.g., sutures or clips), which is also the casewhen bleeding is diffuse.

[0030] In practicing the invention, Factor VIIa or a Factor VIIaequivalent is administered to a patient as a single dose comprising asingle-dose-effective amount. Administration of a single dose refers toadministration of an entire dose of Factor VIIa as a bolus over a periodof less than about 5 minutes. In some embodiments, the administrationoccurs over a period of less than about 2.5 minutes, and, in some, overless than about 1 min.

[0031] A single-dose-effective amount of Factor VIIa or a Factor VIIaequivalent refers to the amount of Factor VIIa or equivalent which, whenadministered in a single dose according to the invention, produces ameasurable improvement in at least one clinical parameter of haemostasisknown to those of ordinary skill in the art (see below). Typically, asingle-dose effective amount comprises at least 150 ug/kg Factor VIIa.In different embodiments, a single-dose-effective amount of Factor VIIacomprises between about 150-500 ug/kg; 250-500 ug/kg; 300-500 ug/kg;350-500 ug/kg; 400-500 ug/kg; 450-500 ug/kg; or more than 500 ug/kg,respectively. When Factor VIIa equivalents are administered according tothe present invention, a single-dose effective amount corresponding tothe above-cited amounts may be determined by comparing the anticoagulantactivity of the Factor VIIa equivalent with that of Factor VIIa (seebelow) and adjusting the amount to be administered proportionately.

[0032] It will be understood that a single-dose-effective amount ofFactor VIIa may vary according to the subject's haemostatic status,which, in turn, may be reflected in one or more clinical parameters,including, e.g., relative levels of circulating coagulation factors;amount of blood lost; rate of bleeding; hematocrit, and the like. Itwill be further understood that the single-dose-effective amount may bedetermined by those of ordinary skill in the art by routineexperimentation, by constructing a matrix of values and testingdifferent points in the matrix.

[0033] In some embodiments, following administration of a single-dose ofFactor VIIa or a Factor VIIa equivalent according to the invention, thepatient receives no further Factor VIIa or Factor VIIa equivalent for aninterval of at least about 1 hour. In some embodiments thepost-administration interval is at least about 4 hours; in otherembodiments, the post-administration interval is at least about 24hours. In still other embodiments, no further Factor VIIa or Factor VIIaequivalent is administered to treat the particular bleeding episode.

[0034] According to the invention, Factor VIIa or a Factor VIIaequivalent may be administered by any effective route, including,without limitation, intravenous, intramuscular, subcutaneous, mucosal,and pulmonary routes of administration. Preferably, administration is byan intravenous route.

[0035] Factor VIIa and Factor VIIa equivalents: In practicing thepresent invention, any Factor VIIa or equivalent may be used that iseffective in preventing or treating bleeding when administered in asingle dose. In some embodiments, the Factor VIIa is human Factor VIIa,as disclosed, e.g., in U.S. Pat. No. 4,784,950 (wild-type Factor VII).The term “Factor VII” is intended to encompass Factor VII polypeptidesin their uncleaved (zymogen) form, as well as those that have beenproteolytically processed to yield their respective bioactive forms,which may be designated Factor VIIa. Typically, Factor VII is cleavedbetween residues 152 and 153 to yield Factor VIIa.

[0036] Factor VIIa equivalents include, without limitation, Factor VIIpolypeptides that have either been chemically modified relative to humanFactor VIIa and/or contain one or more amino acid sequence alterationsrelative to human Factor VIIa. Such equivalents may exhibit differentproperties relative to human Factor VIIa, including stability,phospholipid binding, altered specific activity, and the like.

[0037] In one series of embodiments, a Factor VIIa equivalent includespolypeptides that exhibit at least about 10%, preferably at least about30%, more preferably at least about 50%, and most preferably at leastabout 70%, of the specific biological activity of human Factor VIIa. Forpurposes of the invention, Factor VIIa biological activity may bequantified by measuring the ability of a preparation to promote bloodclotting using Factor VII-deficient plasma and thromboplastin, asdescribed, e.g., in U.S. Pat. No. 5,997,864. In this assay, biologicalactivity is expressed as the reduction in clotting time relative to acontrol sample and is converted to “Factor VII units” by comparison witha pooled human serum standard containing 1 unit/ml Factor VII activity.Alternatively, Factor VIIa biological activity may be quantified by (i)measuring the ability of Factor VIIa or a Factor VIIa equivalent toproduce of Factor Xa in a system comprising TF embedded in a lipidmembrane and Factor X. (Persson et al., J. Biol. Chem. 272:19919-19924,1997); (ii) measuring Factor X hydrolysis in an aqueous system (see,Example 5 below); (iii) measuring the physical binding of Factor VIIa ora Factor VIIa equivalent to TF using an instrument based on surfaceplasmon resonance (Persson, FEBS Letts. 413:359-363, 1997) and (iv)measuring hydrolysis of a synthetic substrate by Factor VIIa and/or aFactor VIIa equivalent.

[0038] Non-limiting examples of Factor VII-related polypeptides havingsubstantially the same or improved biological activity as wild-typeFactor VII include S52A-FVII, S60A-FVII (lino et al., Arch. Biochem.Biophys. 352: 182-192, 1998); L305V-FVII, L305V/M306D/D309S-FVII,L3051-FVII, L305T-FVII, F374P-FVII, V158T/M298-Q-FVII,V158D/E296V/M298Q-FVII, K337A-FVII, M298Q-FVII, V158D/M298Q-FVII,L305V/K337A-FVII, V158D/E296V/M298Q/L305V-FVII,V158D/E296V/M298Q/K337A-FVII, V158D/E296V/M298Q/L305V/K337A-FVII,K157A-FVII, E296V-FVII, E296V/M298Q-FVII, V158D/E296V-FVII,V158D/M298K-FVII, and S336G-FVII; Factor VIIa variants exhibitingincreased proteolytic stability as disclosed in U.S. Pat. No. 5,580,560;Factor VIIa that has been proteolytically cleaved between residues 290and 291 or between residues 315 and 316 (Mollerup et al., Biotechnol.Bioeng. 48:501-505, 1995); oxidized forms of Factor VIIa (Komfelt etal., Arch. Biochem. Biophys. 363:43-54, 1999), Factor VII-sequencevariants wherein the amino acid residue in positions 290 and/or 291 (ofSEQ ID NO:1), preferably 290, have been replaced, and FactorVII-sequence variants wherein the amino acid residue in positions 315and/or 316 (of SEQ ID NO:1), preferably 315, have been replaced.

[0039] Preparations and formulations: The present invention encompassestherapeutic administration of Factor VIIa or Factor VIIa equivalents,which is achieved using formulations that comprise Factor VIIapreparations. As used herein, a “Factor VII preparation” refers to aplurality of Factor VIIa polypeptides or Factor VIIa equivalentpolypeptides, including variants and chemically modified forms, thathave been separated from the cell in which they were synthesized,whether a cell of origin or a recombinant cell that has been programmedto synthesize Factor VIIa or a Factor VIIa equivalent.

[0040] Separation of polypeptides from their cell of origin may beachieved by any method known in the art, including, without limitation,removal of cell culture medium containing the desired product from anadherent cell culture; centrifugation or filtration to removenon-adherent cells; and the like.

[0041] Optionally, Factor VII polypeptides may be further purified.Purification may be achieved using any method known in the art,including, without limitation, affinity chromatography, such as, e.g.,on an anti-Factor VII antibody column (see, e.g., Wakabayashi et al., J.Biol. Chem. 261:11097, 1986; and Thim et al., Biochem. 27:7785, 1988);hydrophobic interaction chromatography; ion-exchange chromatography;size exclusion chromatography; electrophoretic procedures (e.g.,preparative isoelectric focusing (IEF), differential solubility (e.g.,ammonium sulfate precipitation), or extraction and the like. See,generally, Scopes, Protein Purification, Springer-Verlag, New York,1982; and Protein Purification, J.-C. Janson and Lars Ryden, editors,VCH Publishers, New York, 1989. Following purification, the preparationpreferably contains less than about 10% by weight, more preferably lessthan about 5% and most preferably less than about 1%, of non-Factor VIIproteins derived from the host cell.

[0042] Factor VII and Factor VII-related polypeptides may be activatedby proteolytic cleavage, using Factor XIIa or other proteases havingtrypsin-like specificity, such as, e.g., Factor IXa, kallikrein, FactorXa, and thrombin. See, e.g., Osterud et al., Biochem. 11:2853 (1972);Thomas, U.S. Pat. No. 4,456,591; and Hedner et al., J. Clin. Invest.71:1836 (1983). Alternatively, Factor VII may be activated by passing itthrough an ion-exchange chromatography column, such as Mono Q®(Pharmacia) or the like. The resulting activated Factor VII may then beformulated and administered as described below.

[0043] Pharmaceutical compositions or formulations for use in thepresent invention comprise a Factor VIIa preparation in combinationwith, preferably dissolved in, a pharmaceutically acceptable carrier,preferably an aqueous carrier or diluent. A variety of aqueous carriersmay be used, such as water, buffered water, 0.4% saline, 0.3% glycineand the like. The preparations of the invention can also be formulatedinto liposome preparations for delivery or targeting to the sites ofinjury. Liposome preparations are generally described in, e.g., U.S.Pat. Nos. 4,837,028, 4,501,728, and 4,975,282. The compositions may besterilised by conventional, well-known sterilisation techniques. Theresulting aqueous solutions may be packaged for use or filtered underaseptic conditions and lyophilised, the lyophilised preparation beingcombined with a sterile aqueous solution prior to administration.

[0044] The compositions may contain pharmaceutically acceptableauxiliary substances or adjuvants, including, without limitation, pHadjusting and buffering agents and/or tonicity adjusting agents, suchas, for example, sodium acetate, sodium lactate, sodium chloride,potassium chloride, calcium chloride, etc.

[0045] Combinations: The present invention encompasses combinedsingle-dose administration of an additional agent in concert with FactorVIIa or a Factor VIIa equivalent. In some embodiments, the additionalagent comprises a coagulant, including, without limitation, acoagulation factor such as, e.g., Factor VIII, Factor IX, or FactorXIII; or an inhibitor of the fibrinolytic system, such as, e.g.,aprotinin, ε-aminocaproic acid or tranexamic acid. In other embodiments,the additional agent comprises an anticoagulant, including, withoutlimitation, heparin, warfarin, coumarin, and modified Factor VIIpolypeptides, such as, e.g., R152E-Factor VIIa (Wildgoose et al.,Biochem 29:3413-3420, 1990), S344A-Factor VIIa (Kazama et al., J. Biol.Chem. 270:66-72, 1995), FFR-Factor VIIa (Hoist et al., Eur. J. Vasc.Endovasc. Surg. 15:515-520, 1998), Factor VIIa lacking the Gla domain,(Nicolaisen et al., FEBS Letts. 317:245-249, 1993), and chemicallymodified Factor VII polypeptides (U.S. Pat. No. 5,997,864).

[0046] It will be understood that, in embodiments comprising single-doseadministration of combinations of Factor VIIa with other agents, thedosage of Factor VIIa or Factor VIIa equivalent may on its own comprisea single-dose-effective amount. Alternatively, the combination of FactorVIIa or equivalent and the second agent may together comprise asingle-dose-effect amount for preventing or treating bleeding episodes.

[0047] Indications: The present invention encompasses single-doseadministration of Factor VIIa or a Factor VIIa equivalent to any patientwho either anticipates a bleeding episode or who is activelyexperiencing a bleeding episode. Such patients include, withoutlimitation, those suffering from bleeding disorders that are caused byclotting factor deficiencies (e.g. haemophilia A and B, or deficiency ofcoagulation Factors XI or VII); clotting factor inhibitors; defectiveplatelet function; thrombocytopenia; or von Willebrand's disease. Themethods of the invention may also be applied to patients who are aboutto undergo surgery, preferably major surgery, whether or not they sufferfrom a bleeding disorder; as well as trauma patients. Furthermore, anytype of profuse bleeding from the gastrointestinal tract, or anybleeding occurring postoperatively (including that occurring in patientsnot suffering from a bleeding disorder) may benefit from treatmentaccording to the present invention.

[0048] In some embodiments, the invention does not encompassadministration of Factor VIIa or equivalent to patients undergoing minorsurgery. In other embodiments, the invention does not encompassadministration of Factor VIIa or equivalent to patients not sufferingfrom a clotting disorder who had been administered an anticoagulant(such as, e.g., acetocoumerol) within 48 hours prior to Factor VIIaadministration. In other embodiments, the invention does not encompassadministration of Factor VIIa or equivalent to patients not sufferingfrom a clotting disorder who had been administered a Vitamin Kantagonist within 48 hours prior to Factor VIIa administration.

[0049] The present invention also provides the benefit of allowing apatient to selfadminister an effective dose of Factor VIIa or a FactorVIIa equivalent in order to facilitate effective management ofanticipated or current bleeding episodes.

[0050] Many variations of the present invention will suggest themselvesto those skilled in the art in light of the above detailed description.Such obvious variations are within the full intended scope of theinvention.

[0051] All patents, patent applications, and literature referencesreferred to herein are hereby incorporated by reference in theirentirety.

[0052] The following examples are intended as non-limiting illustrationsof the present invention.

EXAMPLE 1

[0053] High-dose Factor VIIa Administration

[0054] Patients suffering from hemophilia (including, e.g., patientswith clotting factor inhibitors, acquired inhibitor patients, patientssuffering from Factor VII deficiency, and patients suffering from vonWillebrands disease) are enrolled in a registry that tracks the outcomeof bleeding episodes whose treatment includes bolus administration ofFactor VIIa.

[0055] The bleeding episodes are characterized as spontaneous, relatedto traumatic injury, or other (including, e.g., surgical or dentalprocedures). The dosage groups are characterized as <100 ug/kg; 100-150ug/kg; 150-200 ug/kg; and >200 ug/kg. Responses to treatment (asassessed at 72 h) are characterized as cessation of bleeding, slowing ofbleeding; or no response.

EXAMPLE 2

[0056] Single High-dose Factor VIIa Administration

[0057] Description of Clinical Trial:

[0058] A randomized, multicenter, cross-over, double-blind study isperformed to evaluate the efficacy and safety of Factor VIIa (by twodifferent blinded dose schedules) in producing hemostasis in jointbleeds in a home-treatment setting. Subjects with congenital hemophiliaA or B and inhibitors to Factor VIII or Factor IX receive treatment andare assessed for at least 9 hours after the dosing. The success orfailure of the treatment is ascertained using a pilot algorithm toassess changes in pain and joint mobility. rFVIIa will be given as anintravenous bolus injection, either at 270 μg/kg body weight dose athour 0, or at 90 μg/kg body weight doses given at hours 0, 3 and 6 andplacebo solutions will be administered to blind subject as to the doseregimen of rFVIIa being administered. If additional doses of Factor VIIaare administered within the first 9 hours to achieve hemostasis, thenthe treatment efficacy is graded as a failure.

[0059] Trial Population:

[0060] Twenty-four patients with congenital hemophilia A or B andinhibitors to factor VIII or IX have been enrolled in this trial.Patients must have experienced two or more mild or moderate joint bleedsduring the past 12 months.

[0061] Assessments

[0062] Treatment efficacy is based on the evaluation of pain, jointmobility and measure of circumference of the elbow or knee at themidpoint of the joint in extension. These variables are graded andentered in the diary by the patient/caregiver. Pain and mobility areassessed as more, no difference or less than before the treatment andcircumference is measured in millimeters. An independent blindedcommittee reviews the diary data on pain and joint mobility and judgesthe response to treatment as success or failure based on a pilotalgorithm developed for this study.

1. A method for treating a bleeding episode in a subject in need of suchtreatment, said method comprising administering to said subject FactorVIIa or a Factor VIIa equivalent, wherein said administering is in asingle dose and said dose comprises a single-dose-effective amount ofsaid Factor VIIa or Factor VIIa equivalent.
 2. A method as defined inclaim 1, wherein, subsequent to said administration, no further FactorVIIa or protein having Factor VIIa coagulant activity is administered tosaid subject for a period of at least about 1 hour.
 3. A method asdefined in claim 2, wherein said period is at least about 4 hours.
 4. Amethod as defined in claim 3, wherein said period is at least about 24hours.
 5. A method as defined in claim 1, wherein said single dose isadministered over a period of less than about 5 minutes.
 6. A method asdefined in claim 1, wherein said single-dose-effective amount comprisesbetween about 150 and about 500 ug/kg Factor VIIa or a correspondingamount of a Factor VIIa equivalent.
 7. A method as defined in claim 6,wherein said single-dose-effective amount comprises between about 200and about 500 ug/kg Factor VIIa or a corresponding amount of a FactorVIIa equivalent.
 8. A method as defined in claim 7, wherein saidsingle-dose-effective amount comprises between about 250 and about 500ug/kg Factor VIIa or Factor VIIa equivalent or a corresponding amount ofa Factor VIIa equivalent.
 9. A method as defined in claim 8, whereinsaid single-dose-effective amount comprises between about 300 and about500 ug/kg Factor VIIa or a corresponding amount of a Factor VIIaequivalent.
 10. A method as defined in claim 1, wherein said Factor VIIaequivalent exhibits at least about 30% of the coagulant activity ofFactor VIIa on a molar basis.
 11. A method as defined in claim 1,wherein said Factor VIIa equivalent is selected from the groupconsisting of: S52A-FVII, S60A-FVII; L305V-FVII, L305V/M306D/D309S-FVII,L3051-FVII, L305T-FVII, F374P-FVII, V158T/M298-Q-FVII,V158D/E296V/M298Q-FVII, K337A-FVII, M298Q-FVII, V158D/M298Q-FVII,L305V/K337A-FVII, V158D/E296V/M298Q/L305V-FVII,V158D/E296V/M298Q/K337A-FVII, V158D/E296V/M298Q/L305V/K337A-FVII,K157A-FVII, E296V-FVII, E296V/M298Q-FVII, V158D/E296V-FVII,V158D/M298K-FVII, and S336G-FVII; Factor VIIa that has beenproteolytically cleaved between residues 290 and 291; Factor VIIa thathas been proteolytically cleaved between residues 315 and 316; andoxidized forms of Factor VIIa.
 12. A method as defined in claim 1,wherein said bleeding episode comprises a major bleed.
 13. A method asdefined in claim 1, further comprising administering, in said singledose, or substantially simultaneously with said single dose, a secondcoagulant agent.
 14. A method as defined in claim 12, wherein saidsecond coagulant agent is selected from the group consisting of FactorVIII, Factor IX, and Factor XIII.
 15. A method as defined in claim 1,further comprising administering an anticoagulant, wherein saidanticoagulant is administered in said single dose, or substantiallysimultaneously with said single dose.
 16. A method for treating ableeding episode, said method comprising administering to a subject inneed of such treatment (i) a first amount of Factor VIIa or a FactorVIIa equivalent and (ii) a second amount of second coagulant agent,wherein said first and second amounts together comprise an aggregateeffective amount for treating said bleeding and said aggregate effectiveamount is administered in a single dose.
 17. A method as defined inclaim 1, wherein said Factor VIIa is recombinant human Factor VIIa. 18.A method for treating a bleeding episode, said method comprisingadministering to a human subject in need of such treatment an effectiveamount for treating said bleeding of human Factor VIIa or a human FactorVIIa equivalent, wherein: (i) said effective amount is administered in asingle dose over a period of less than about 5 minutes; (ii) saideffective amount comprises between about 300 and about 500 ug/kg humanFactor VIIa or human Factor VIIa equivalent or a corresponding amount ofa Factor VIIa equivalent; and (iii) subsequent to said administration,no further Factor VIIa or Factor VIIa equivalent is administered to saidsubject for a period of at least about 1 hour.
 19. A method as definedin claim 1, wherein said administering is via a route selected from thegroup consisting of intravenous, intramuscular, subcutaneous, mucosal,and pulmonary administration.
 20. A method for preventing bleeding, saidmethod comprising administering to said subject Factor VIIa or a FactorVIIa equivalent, wherein said administering is in a single dose and saiddose comprises a single-dose-effective amount of said Factor VIIa orFactor VIIa equivalent.
 21. A method as defined in claim 20, wherein,subsequent to said administration, no further Factor VIIa or proteinhaving Factor VIIa coagulant activity is administered to said subjectfor a period of at least about 1 hour.
 22. A method as defined in claim21, wherein said period is at least about 4 hours.
 23. A method asdefined in claim 22, wherein said period is at least about 24 hours. 24.A method as defined in claim 20, wherein said single dose isadministered over a period of less than about 5 minutes.
 25. A method asdefined in claim 20, wherein said single-dose-effective amount comprisesbetween about 150 and about 500 ug/kg Factor VIIa or a correspondingamount of a Factor VIIa equivalent.
 26. A method as defined in claim 25,wherein said single-dose-effective amount comprises between about 200and about 500 ug/kg Factor VIIa or a corresponding amount of a FactorVIIa equivalent.
 27. A method as defined in claim 26, wherein saidsingle-dose-effective amount comprises between about 250 and about 500ug/kg Factor VIIa or Factor VIIa equivalent or a corresponding amount ofa Factor VIIa equivalent.
 28. A method as defined in claim 27, whereinsaid single-dose-effective amount comprises between about 300 and about500 ug/kg Factor VIIa or a corresponding amount of a Factor VIIaequivalent.
 29. A method as defined in claim 20, wherein said FactorVIIa equivalent exhibits at least about 30% of the coagulant activity ofFactor VIIa on a molar basis.
 30. A method as defined in claim 20,wherein said Factor VIIa equivalent is selected from the groupconsisting of: S52A-FVII, S60A-FVII; L305V-FVII, L305V/M306D/D309S-FVII,L3051-FVII, L305T-FVII, F374P-FVII, V158T/M298-Q-FVII,V158D/E296V/M298Q-FVII, K337A-FVII, M298Q-FVII, V158D/M298Q-FVII,L305V/K337A-FVII, V158D/E296V/M298Q/L305V-FVII,V158D/E296V/M298Q/K337A-FVII, V158D/E296V/M298Q/L305V/K337A-FVII,K157A-FVII, E296V-FVII, E296V/M298Q-FVII, V158D/E296V-FVII,V158D/M298K-FVII, and S336G-FVII; Factor VIIa that has beenproteolytically cleaved between residues 290 and 291; Factor VIIa thathas been proteolytically cleaved between residues 315 and 316; andoxidized forms of Factor VIIa.
 31. A method as defined in claim 20,wherein said bleeding episode comprises a major bleed.
 32. A method asdefined in claim 20, further comprising administering, in said singledose, or substantially simultaneously with said single dose, a secondcoagulant agent.
 33. A method as defined in claim 32, wherein saidsecond coagulant agent is selected from the group consisting of FactorVIII, Factor IX, and Factor XIII.
 34. A method as defined in claim 20,further comprising administering an anticoagulant, wherein saidanticoagulant is administered in said single dose, or substantiallysimultaneously with said single dose.
 35. A method for treating ableeding episode, said method comprising administering to a subject inneed of such treatment (i) a first amount of Factor VIIa or a FactorVIIa equivalent and (ii) a second amount of second coagulant agent,wherein said first and second amounts together comprise an aggregateeffective amount for treating said bleeding and said aggregate effectiveamount is administered in a single dose.
 36. A method as defined inclaim 20, wherein said Factor VIIa is recombinant human Factor VIIa. 37.A method for preventing a bleeding episode, said method comprisingadministering to a human subject in need of such treatment an effectiveamount for treating said bleeding of human Factor VIIa or a human FactorVIIa equivalent, wherein: (i) said effective amount is administered in asingle dose over a period of less than about 5 minutes; (ii) saideffective amount comprises between about 300 and about 500 ug/kg humanFactor VIIa or human Factor VIIa equivalent or a corresponding amount ofa Factor VIIa equivalent; and (iii) subsequent to said administration,no further Factor VIIa or Factor VIIa equivalent is administered to saidsubject for a period of at least about 1 hour.
 38. A method as definedin claim 20, wherein said administering is via a route selected from thegroup consisting of intravenous, intramuscular, subcutaneous, mucosal,and pulmonary administration.